Urine trypsin inhibitors which are a kind of enzymatic activity inhibitor contained in urines of mammalians are known to vary in characteristic properties depending upon the species of animals from which they are derived [Carlson, Enzyme, 18, 176 (1974)]. HUTI, a glucoprotein having a molecular weight of 68,000 and an isoelectric point of pH 2 to 3, inhibits particularly trypsin, chymotrypsin, plasmin [Sumi, et al., Journal of Physiol. of Japan, 39, 53 (1977)], human esterase [Johnson et al., Hoppe Seyler's Z. Physiol. Chem., 363, 1167 (1982)], etc., and is possibly considered to have application as an anti-inflammatory agent. Kallidinogenase is a proteolytic enzyme produced in every part of the living bodies of animals, and, through its decomposition of kininogen in serum to produce kinin, demonstrates diverse activities, such as blood pressure decreasing and blood flow increasing activities. Both HUTI and HUKN have previously been separated from different species of animals than man, and attempts have been made to utilize them as drugs. Especially, kallidinogenase, which has been produced by using porcine pancreas and urine as raw materials. However, these are proteins foreign to man and exhibit antigenicity. When they are used clinically, frequent administration often produces anaphylaxis, and the problem has been encountered from the standpoint of safety. In contrast to this, HUTI and HUKN are proteins of human origin, and consequently, are entirely free from the aforementioned side effects in humans.
Conventional concentration purification methods for HUTI include, for example, the precipitation method involving the combined use of trichloroacetic acid, ammonium sulfate and methanol [N. R. Shulman: J. B. C., 213, 655 (1955)], the method utilizing bentonite [T. A. Strup: Scan. J. Clin. Lab. Invest., 11, 181 (1959)] and the method using DEAE-cellulose [G. J. Proksch: J. Lab & Clin. Med., 79, 491 (1972)]. With reference to the conventional concentration purification method for HUKN, there are known, for example, the method using silica gel (Japanese Patent Application laid-open No. 99151/1980) and the method utilizing a microporous type of ion exchange resin, such as WA-30 (Japanese Patent Application laid-open No. 5515/1975) as well as the concentration purification method for HUKN developed by the present inventors (Japanese Patent Application laid-open No. 6883/1984) which consists of using chitosan as an adsorbent. A known method of concentrating simultaneously both HUTI and HUKN, is the method of Sumi et al. [Sumi, et al.: Medicine and Biology ("Igaku to Seibutsugaku", vol. 100 (1) (1980)] which involves the use of Arginine-Sepharose. Referring to the above-mentioned methods, those intended for use in the purification and processing into preparations of either HUTI or HUKN alone are not always satisfactory in terms of cost incurred and operation involved. The method of concentrating simultaneously both of these requires too much expensive adsorbent and requires too complex an operational procedure to be practiced on an industrial scale.